QUANTITATIVE DETECTION OF HEPATITIS-C VIRUS-RNA BY POLYMERASE CHAIN-REACTION AND DOT-BLOT HYBRIDIZATION

We have developed a new technique to quantify hepatitis C virus (HCV)- RNA using polymerase chain reaction (PCR) and dot blot hybridization. PCR was performed on 10-fold dilutions of an HCV-cDNA fragment for var ious numbers of cycles and on various serum volumes for 30 cycles. The resulting PCR products were spotted onto nitrocellulose filters and h ybridized with a P-32-labelled internal probe. The intensity of the hy bridization signal was proportional to the quantity of template cDNA a nd increased in proportion to the number of PCR cycles. Clear signals were detected after 30 cycles of PCR using an original serum volume of greater than 250 mul. Thus, this new technique allows the quantitatio n of HCV-RNA, and as it is simple and cost-effective, it promises to h ave many clinical applications.